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SCIATIC NERVE REGENERATION USING CALCIUM PHOSPHATE COATED CONDUIT AND BRAIN-DERIVED NEUROTROPHIC FACTOR GENE-TRANSFECTED SCHWANN CELL IN RAT (인회석 박막 피복 도관과 Brain-derived neurotrophic factor(BDNF) 유전자 이입 슈반세포를 이용한 백서 좌골신경 재생에 관한 연구)

  • Choi, Won-Jae;Ahn, Kang-Min;Hwang, Soon-Jeong;Choung, Pill-Hoon;Kim, Myung-Jin;Kim, Nam-Yeol;Yoo, Sang-Bae;Jahng, Jeong-Won;Kim, Hyun-Man;Kim, Joong-Soo;Kim, Yun-Hee;Kim, Soung-Min;Lee, Jong-Ho
    • Journal of the Korean Association of Oral and Maxillofacial Surgeons
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    • v.31 no.3
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    • pp.199-218
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    • 2005
  • Purpose of Study: Peripheral nerve regeneration depends on neurotrophism of distal nerve stump, recovery potential of neuron, supporting cell like Schwann cell and neurotrophic factors such as BDNF. Peripheral nerve regeneration can be enhanced by the conduit which connects the both sides of transected nerve. The conduit maintains the effects of neurotrophism and BDNF produced by Schwann cells which can be made by gene therapy. In this study, we tried to enhance the peripheral nerve regeneration by using calcium phosphate coated porous conduit and BDNF-Adenovirus infected Schwann cells in sciatic nerve of rats. Materials and Methods: Microporous filter which permits the tissue fluid essential for nerve regeneration and does not permit infiltration of fibroblasts, was made into 2mm diameter and 17mm length conduit. Then it was coated with calcium phosphate to improve the Schwann cell adhesion and survival. The coated filter was evaluated by SEM examination and MTT assay. For effective allogenic Schwann cell culture, dorsal root ganglia of 1-day old rat were extracted and treated with enzyme and antimitotic Ara-C. Human BDNF cDNA was obtained from cDNA library and amplified using PCR. BDNF gene was inserted into adenovirus shuttle vector pAACCMVpARS in which E1 was deleted. We infected the BDNF-Ad into 293 human mammary kidney cell-line and obtained the virus plaque 2 days later. RT-PCR was performed to evaluate the secretion of BDNF in infected Schwann cells. To determine the most optimal m.o.i of BDNF-Ad, we infected the Schwann cells with LacZ adenovirus in 1, 20, 50, 75, 100, 250 m.o.i for 2 hours and stained with ${\beta}$-galactosidase. Rats(n=24) weighing around 300g were used. Total 14mm sciatic nerve defect was made and connected with calcium phosphate coated conduits. Schwann cells$(1{\times}10^6)$ or BDNF-Ad infected Schwann cells$(1{\times}10^6)$ were injected in conduit and only media(MEM) was injected in control group. Twelve weeks after surgery, degree of nerve regeneration was evaluated with gait analysis, electrophysiologic measurements and histomorphometric analysis. Results: 1. Microporous Millipore filter was effective conduit which permitted the adhesion of Schwann cells and inhibited the adhesion of fibroblast. We could enhance the Schwann cell adhesion and survival by coating Millipore filter with calcium phosphate. 2. Schwann cell culture technique using repeated treatment of Ara-C and GDNF was established. The mean number of Schwann cells obtained 1 and 2 weeks after the culture were $1.54{\pm}4.0{\times}10^6$ and $9.66{\pm}9.6{\times}10^6$. 3. The mRNA of BDNF in BDNF-Ad infected Schwann cells was detected using RT-PCR. In Schwann cell $0.69\;{\mu}g/{\mu}l$ of DNA was detected and in BDNF-Adenovirus transfected Schwann cell $0.795\;{\mu}g/{\mu}l$ of DNA was detected. The most effective infection concentration was determined by LacZ Adenovirus and 75 m.o.i was found the most optimal. Conclusion: BDNF-Ad transfected Schwann cells successfully regenerated the 14mm nerve gap which was connected with calcium phosphate coated Millipore filter. The BDNF-Ad group showed better results compared with Schwann cells only group and control group in aspect to sciatic function index, electrophysiologic measurements and histomorphometric analysis.

The Effects of Prostaglandin and Dibutyryl cAMP on Osteoblastic Cell Activity and Osteoclast Generation (Prostaglandin과 Dibutyryl cAMP가 조골세포의 활성과 파골세포 형성에 미치는 영향)

  • Mok, Sung-Kyu;You, Hyung-Keun;Shin, Hyung-Shik
    • Journal of Periodontal and Implant Science
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    • v.26 no.2
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    • pp.448-468
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    • 1996
  • To maintain its functional integrity, bone is continuously remodelled by a process involving resorption by osteoeclasts and formation by osteoblasts, In order to respond to changes in the physical environment or to trauma with the relevant action, this process is strictly regulated by locally synthesized or systemic fators, Prostaglandin $E_2(PGE_2$) is perhaps one of the best studied factors, having been known to affect bone cell function for several decades.$PGE_2$ has both anabolic and catabolic activities. Excess of $PGE_2$ has been implicated in a number of pathological states associated with bone loss in a number of chronic inflammatory conditions such as periodontal disease and rheumatoid arthritis. $PGE_2$ and other arachidonic acid metabolites have been shown to be potent stimulators of osteoclastic bone resorption in organ culture. The anabolic effects of $PGE_2$ were first noticed when an increase in periosteal woven bone formation was seen after the infusion of $PGE_2$ into infants in order to prevent closure of the ductus arteriosus. The cellular basis for the catabolic actions of $PGE_2$ has been well characterized. $PGE_2$increases osteoclast recruitment in bone marrow cell cultures. Also $PGE_2$ has a direct action on osteoclast serving to inhibit activity and can also indirectly activate osteoclast via other cells in the vicinity, presumably osteoblast. The cellular mechanisms for the anabolic actions of $PGE_2$ are not nearly so well understood. The purpose of this paper was to study the effects of $PGE_2$ and dibutyl(DB)cAMP on osteoblastic clone MC3T3El cells and on the generation of osteoclasts from their precursor cells. The effect of $PGE_2$ and DBcAMP on the induction of alkaline phoaphatase(AlP) was investigated in osteoblastic clone MC3T3El cells cultured in medium containing 0.4% fetal bovine serum. $PGE_2$ and DBcAMP stimulated ALP activity and MTT assay in the cells in a dose-dependent manner at concentrations of lO-SOOng/ml. Cycloheximide, protein synthesis inhibitor, inhibited the stimulative effect of $PGE_2$ and DBcAMP on ALP activity in the cells. $PGE_2$also increased the intracellular cAMP content in a dose-dependent fashion with a maximal effect at 500ng/ml. The effect of $PGE_2$ on the generation of osteoclasts was investigated in a coculture system of mouse bone marrow cells with primary osteoblastic cells cultured in media containing 10% fetal bovine serum.After cultures, staining for tartrate-resistant acid phosphatase(TRAP)-marker enzyme of osteoclast was performed. The TRAP(+) multinucleated cells(MNCs), which have 3 or more nuclei, were counted. More TRAP(+) MNCs were formed in coculture system than in control group. $PGE_2(10^{-5}10^{-6}M)$ stimulated the formation of osteoclast cells from mouse bone marrow cells in culture. $PGE_2(10^{-6}M)$ stimulated the formation of osteoclast cells from mouse bone marrow cells in coculture of osteoblastic clone MC3T3E1 cells This results suggest that $PGE_2$ stimulates the differentiation of osteoblasts and generation of osteoclast, and are involved in bone formation, as well as in bone resorption.

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Intestinal Atresia - The Second National Survey (선천성 장폐쇄증 2차 조사 - 대한소아외과학회 회원을 주 대상으로 한 전국조사 -)

  • Kim, Seong-Chul;Kim, D.Y.;Kim, S.Y.;Kim, I.K.;Kim, I.S.;Kim, J.E.;Kim, J.C.;Kim, H.Y.;Park, K.W.;Park, W.H.;Park, J.Y.;Seo, J.M.;Seol, J.Y.;Oh, S.M.;Oh, J.T.;Lee, N.H.;Lee, M.D.;Lee, S.K.;Lee, S.C.;Chung, S.Y.;Jung, S.E.;Jeong, Y.J.;Jung, P.M.;Choi, K.J.
    • Advances in pediatric surgery
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    • v.16 no.1
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    • pp.1-10
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    • 2010
  • The members of the Korean Association of Pediatric Surgeons conducted a retrospective study of two hundred and twenty-two cases of intestinal atresia for the period from January 1, 2007 to December 31, 2009. Seventeen hospitals were involved. There were 76 duodenal, 65 jejunal, and 81 ileal atresias (3 colonic). The male to female ratio was 0.85:1 in DA and 1.34:1 in JIA. Ninety-four patients(43.3 %) were premature babies (DA 40.3 %, JA 64.6 %, IA 28.8 %), and 70 babies (32.0 %) had low birth weight (DA 38.7 %, JA 44.4 %, IA 16.0 %). Antenatal diagnosis was made in 153 cases (68.9 %). However, 27 infants (17.6 %) with antenatal diagnosis were transferred to the pediatric surgeon's hospitals after delivery. Maternal polyhydramnios was observed in 81 cases (36.59 %) and most frequent with proximal obstruction. In forty-four cases (19.8 %), only simple abdominal film was taken for diagnostic study. The associated malformations were more frequently observed in DA - 61.8 % in DA and 22.6 % in JIA. Meconium peritonitis, small bowel volvulus and intussusception were more frequently associated with ileal atresia. The overall mortality rate was 3.6 %. (Abbreviations: DA;duodenal atersia, JA;jejunal atresia, JIA;jejunoileal atresia, IA;ileal atrsia).

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Comparison of PCR-Line Probe and PCR-SSCP Methods for the Detection of Rifampicin Resistant Mycobacterium Tuberculosis (Rifampicin 내성 결핵균의 검출에 있어서 PCR-line Probe법과 PCR-SSCP법의 비교)

  • Kim, Ho-Joong;Suh, Gee-Young;Chung, Man-Pyo;Kim, Jong-Won;Shim, Tae-Sun;Choi, Dong-Chull;Kwon, O-Jung;Rhee, Chong-H;Han, Yong-Chol
    • Tuberculosis and Respiratory Diseases
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    • v.45 no.4
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    • pp.714-722
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    • 1998
  • Background: Rifampicin (RFP) is a key component of the antituberculous short-course chemotherapy and the RFP resistance is a marker of multi-drug resistant (MDR) tuberculosis. RPoB gene encodes the $\beta$-subunit of RNA polymerase of M. tuberculosis which is the target of RFP. And the mutations of rpoB gene have been found in about 96% of rifampicin resistant clinical isolates of M. tuberculosis. So in order to find a rapid and clinically useful diagnostic method in identifying the RFP resistance, we compared the PCR -line probe method with PCR-SSCP for the detection of the rpoB gene mutation in cultured M. tuberculosis. Methods: 45 clinical isolates were collected from patients who visited Sung Kyun Kwan University Hospital. The RFP susceptibility test was referred to the referral laboratory of the Korean Tuberculosis Institute. 33 were rifampicin resistant and 12 were rifampicin susceptible. The susceptibility results were compared with the results of the PCR-BSCP and PCR-line probe method. Results: We could find rpoB mutations in 27/33(81.8%) RFP-resistant strains by PCR-line probe method, and in 23/33 (69.7%) by PCR-SSCP and there was no significant difference between two methods. There was no mutation in rifampicinn susceptible strains by both methods. Conclusion: PCR-line probe method would be a rapid, sensitive and specific method for the detection of rifampicin resistant Mycobacterium tuberculosis.

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The effect of 8 French catheter and chest tube on the treatment of spontaneous pneumothorax (자연기흉에 있어서 8 French 도관과 흉관의 삽입 치료 효과)

  • Kang, Yoon-Jeong;Koh, Hyoung-Gee;Shin, Jong-Wook;Lim, Seong-Yong;Choi, Jae-Sun;Yu, Ji-Hoon;Park, In-Won;Choi, Byoung-Whui;Hue, Sung-Ho;Seo, Seung-Cheon
    • Tuberculosis and Respiratory Diseases
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    • v.43 no.3
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    • pp.410-419
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    • 1996
  • Background : Spontaneous pneumothorax have been managed with a variety of methods. The technique most frequently used is chest tube drainage. Small caliber catheters were first used in the management of pneumothorax complicating the percutaneous needle aspiration lung biopsy, and the try to treat spontaneous pneumothorax also has been reported. However, the value of small caliber catheters in spontaneous pneumothorax has not been fully evaluated. So, we tried to elucidate the efficacy of 8 French catheter in the management of spontaneous pneumothorax. Method : From January, 1990, to April, 1994, 44 patients with spontaneous pneumothorax treated at Chung-Ang university hospital were reviewed. The patients were sub-divide into 8 French catheter insertion group (n=21) and chest tube insertion group (n=23). We compared the presence of underlying lung disease, the extent of the collapse, the duration of indwelling catheter and complication between two groups. Results : 1) The duration of indwelling showed no significant difference between 8 French catheter group and chest tube. But, complication after insertion as subcutaneous emphysema was developed in only chest tube group. (p<0.05) 2) In the primary spontaneous pneumothorax, all case of the pneumothorax of which size was less than 50% showed complete healing with 8 French catheter insertion. Whereas the success rate in patients with large pneumothorax (more than 50%) was tended to be dependent on the age. 3) In the patients with secondary spontaneous pneumothorax who were managed with 8 French catheter, the success rate was trended to be high if the underlying disease of pneumothorax was not COPD and if the patient was young. Conclusion : These results show that 8 French catheter insertion probably was effective in the pneumothorax less than 50%, the primary spontaneous pneumothorax, young age or secondary pneumothorax not associated with COPD.

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Adherence-induced gene expression in human alveolar macrophages (표면부착에 의한 사람 폐포대식세포의 유전자 발현에 관한 연구)

  • Chung, Man Pyo;Yoo, Chul Gyu;Han, Sung Koo;Shim, Young-Soo;Rhee, Chong H.;Han, Yang Chol;Kim, Young Whan
    • Tuberculosis and Respiratory Diseases
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    • v.43 no.6
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    • pp.936-944
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    • 1996
  • Background: Neutrophils or monocytes separated in vitro by the adherence to plastic surface are known to be activated by surface adherence itself and subsequent experimental data might be altered by surface adherence. Adhesion molecules and gene transcription of the inflammatory mediators are known to be associated in this process. To evaluate whether adhesion molecule and transcriptional activation of the inflammatory substances are also involved in the activation of human alveolar macrophage by the adherence procedure, we designed this experiment. Method : Bronchoalveolar lavage was performed in the person whose lung of either side was confirmed to be nonnal by chest cr and alveolar macrophage was harvested. To measure the expression of Interleukin-8(IL-8) mRNA, manganese superoxide dismutase(SOD) mRNA and CD11/CD18 mRNA in human alveolar macrophage of both adherence state and suspension state, Northern blot analysis was done at 0, 2, 4, 8 and 24hrs after the adherence to plastic surface and during suspension state. Then, phorbol myristate acetate(pMA) and N-formyl-methionyl-leucyl-phenylalanine(fMLP) were added respectively in the same experimental condition. Result : 1) Human alveolar macrophages in the adherent state induced IL-8 mRNA and SOD mRNA expression which was maximal at 8 hours after the adherence to plastic surface. But we could not observe the upregulation of CD18 mRNA by surface adherence. 2) PMA induced these mRNA expression both in the adherent cell and the nonadherem cells, but the induction of mRNA expression by fMLP occurred only in the adherent cells. Conclusion: These results suggest that adherence of huamn alveolar macropahge is an important cell-activating event that may play a critical role in the modulation of lung inflammatory respones.

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Food Component Characteristics of Red-tanner Crab (Chionoecetes japonicus) Paste as Food Processing Source (식품가공소재로서 게 페이스트의 식품성분 특성)

  • Kim, Hye-Suk;Park, Chan-Ho;Choi, Seung-Geal;Han, Byung-Wook;Kang, Kyung-Tae;Shim, Nam-Hyuk;Oh, Hyeon-Seok;Kim, Jin-Soo;Heu, Min-Soo
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.34 no.7
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    • pp.1077-1081
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    • 2005
  • The objective of this study was to investigate the components and nutritional quality of red-tanner crab (Chionoecetes japonicus) paste in order to explore possibility for food processing source such as surimi gel containing crab paste. Yield of crab paste was $30\%$ from whole body after crushing and dehydrating. Crude protein contents $(9.5\%)$ of crab paste was lower than that $(13.1\%)$ of crab muscle, but fat $(0.5\%)$ and ash contents $(8.0\%)$ of paste were higher than $0.2\%\;and\;1.3\%$ of crab muscle, respectively. Volatile basic nitrogen (VBN) content of the crab paste was lower than those of the edible parts. Total amino acid content (9,497mg/l00g) of paste was lower than that (12,980mg/100g) of muscle. Aspartic acid, glutamic acid, lysine and leucine were the predominant amino acids in the protein fraction. The calcium content (6,539mg/l00g) was higher than those of phosphorus (579mg/100g), and potassium (793mg/100g) while manganese and iron were present in trace amounts. Major fatty acids of total lipid were 16:0, 18:1n-9, 20:5n-3 and 22:6n-3, and no difference of composition between paste and muscle. Sensory evaluation showed that scores of color and flavor of $15\%$ substituted surimi gel increased significantly when compared to surimi gel without crab paste (p<0.05). From the above results, the addition of crab paste enhanced nutrition and functionality of surimi gel.

Trace Metal Contents in Seaweeds from Korean Coastal Area (한국 연안 해조류의 미량금속 함량)

  • Kim, Ji-Hoe;Mok, Jong-Soo;Park, Hee-Yeon
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.34 no.7
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    • pp.1041-1051
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    • 2005
  • In order to assure the safety of the seaweeds, we measured the contents of the trace metals in the seaweeds harvested from Korean coastal area. We collected 620 marine algal samples from fourteen areas, and then analyzed the trace metals such as Hg, Cd, Cr, Cu, Mn, Ni, Pb and Zn. The contents of Hg, Cd, Ni and Mn were higher in the seaweeds collected from Chungnam Taean area. The contents of Cr and Cu were higher in the seaweeds collected from Yeongdeok and Tongyeong area, respectively. And Pb and Zn were highly detected from the seaweeds of Ulsan area. The mean levels of trace metals were high in the order of Zn $(48.02\pm41.20{\mu}g/g)\;>\;Mn (34.63\pm38.95{\mu}g/g)\;>\;Cu(6.29\pm6.52{\mu}g/g)\;>\;Ni(2.17\pm4.00{\mu}g/g)\; >\;Cr(1.61\pm2.35{\mu}g/g)\;>\;Pb(1.28\pm2.37{\mu}g/g)\;>\; Cd(0.67\pm0.75{\mu}g/g)\;>\;Hg(0.02\pm0.02{\mu}g/g)$. Zn was consistently most abundant metal in all samples, followed by Mn and Cu, which are very important for human health. There were higher contents of Hg and Cd in brown algae, the contents of Cr, Cu, Mn, Ni and Pb in green algae, the content of Zn in red algae among them. Additionally, significant linear correlations were observed between a total 28 different pairs of metals, of them, the correlation factor between Cr and Ni showed the highest (r=0.374, p<0.001). The trace metal contents of seaweeds varied with habitats and kinds of samples, nevertheless they showed very safe level in edible seaweeds tested. Scytosiphon lomentaria, which has been known as a little use of food, have shown a clear selectivity for Cr and Pb. Among non-edible seaweeds, Sargassum thunbergii for both Cd and Mn, Lomentaria hakodatensis for Ni and Grateloupia prolongata for Zn have shown a clear selectivity, respectively. Therefore, these seaweeds could be useful as bioindicator for each trace metal pollution.

Effect of the Ethanol Extract of Cassia tora L. on Antioxidative Compounds and Lipid metabolism in Hepatotoxicity of Rats-induced by Ethanol (결명자 에탄올추출물이 알코올 투여 흰쥐의 항산화물질 및 지질대사에 미치는 영향)

  • 최현숙;차선숙;나명순;신길만;이명렬
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.30 no.6
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    • pp.1177-1183
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    • 2001
  • This study was done to investigate the effects of ethanol extract of Cassia semen (Cassia tora L.) on the activities of hepatic oxygen free radicals metabolizing enzymes and blood lipid profile in rats of hepatotoxicity induced by ethanol. Sprague-Dawley male rats weighing 100~160 g were divides into 5 groups; control grouts (CON), Cassia semen ethanol extracts (200 mg/kg) treated group (CEL), ethanol (10 mL/kg, 35%) treated group (ETH), Cassia semen ethanol extracts (200 mg/kg) and ethanol treated group (CE1 ) and Cassia semen ethanol extracts (400 mg/kg ) and ethanol treated group (CE2), respectively. Compared with ETH, the growth rate of CE1 and CE2 were to be increased tendency, and in blood levels of total cholesterol, LDL-cholesterol and the activities of alanine aminotranferase and asparate aminotranferase elevated by ethanol were significantly decreased (p<0.05). It was observed that the activities of superoxide dismutase, catalase, xanthine oxidase and glutathione peroxidase of rat liver increased by ethanol, were more decreased by the treatment of Cassia semen ethanol extract than the only ethanol-treated group. The content of glutathione depleted by ethanol treatment was increased in CE1 and CE2. TBA-reactants of liver increased by ethanol were decreased in CE1 and CE2, compared with ethanol-treated group. These results suggested that ethanol extract of Cassia semen may influence upon the ability of oxygen free radical detoxication and lowering of blood lipid level on ethanol-induced hepatotoxicity in rat.

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Preparation and Quality Characteristics of Seasoned and Dried Fish Slice Products Using Rainbow Trout (Onchorhynchus mykiss) (송어 육포의 제조와 품질 특성)

  • Heu, Min-Soo;Kim, Hyung-Jun;Ham, Joon-Sik;Park, Shin-Ho;Kim, Hye-Suk;Kang, Kyung-Tae;Jee, Seung-Joon;Lee, Jae-Hyoung;Kim, Jin-Soo
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.37 no.3
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    • pp.348-356
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    • 2008
  • For the effective use of rainbow trout, the seasoned and dried rainbow trout slice (SR) was prepared and its characterization was compared with the commercial skipjack tuna (CSS) and pork products (CSP) by determining chemical components and sensory evaluation. The moisture content of SR, 20.5%, was lower, and the lipid content of SR, 7.5%, was higher than those of commercial products, CSS and CSP. The protein content of SR (41.4%) was higher than that of CSS (28.6%), but lower than that of CSP (50.1%). The water activity of SR was 0.654, which was lower than those of CSS and CSP, 0.724 and 0.771, respectively. According to the results, the color of SR was lower in lightness and redness and higher in browning index and ${\Delta}$E value than that of commercial products. The texture of SR was harder than that of CSS, but softer than that of CSP. According to the result of taste value, the taste of SR was stronger than that of CSP, while weaker than that of CSS. The mineral (Ca, P, and Fe), total amino acid contents, and the n-3/n-6 of SR showed significant levels in nutrition and health functional senses.